Stability and integrity of self-assembled bovine parvovirus virus­like particles (BPV­VLPs) of VP2 and combination of VP1VP2 assisted by baculovirus-insect cell expression: a potential logistical platform for vaccine deployment.

BACKGROUND: Bovine parvovirus (BPV) is an autonomous DNA virus with a smaller molecular size and subtle differences in its structural proteins, unlike other animal parvoviruses. More importantly, this virus has the potential to produce visible to silent economic catastrophes in the livestock business, despite receiving very little attention. Parvoviral virus-like particles (VLPs) as vaccines and as logistical platforms for vaccine deployment are well studied. However, no single experimental report on the role of VP1 in the assembly and stability of BPV-VLPs is available. Furthermore, the self-assembly, integrity and stability of the VLPs of recombinant BPV VP2 in comparison to VP1 VP2 Cap proteins using any expression method has not been studied previously. In this study, we experimentally evaluated the self-assembling ability with which BPV virus-like particles (VLPs) could be synthesized from a single structural protein (VP2) and by integrating both VP2 and VP1 amino acid sequences. METHODS: In silico and experimental cloning methods were carried out. His-tagged and without-His-tag VP2 and V1VP2-encoding amino acid sequences were cloned and inserted into pFastbacdual, and insect cell-generated recombinant protein was evaluated by SDS‒PAGE and western blot. Period of infectivity and expression level were determined by IFA. The integrity and stability of the BPV VLPs were evaluated by transmission electron microscopy. The secondary structure of the BPV VLPs from both VP2 and V1VP2 was analyzed by circular dichroism. RESULTS: Our findings show that VP2 alone was equally expressed and purified into detectable proteins, and the stability at different temperatures and pH values was not appreciably different between the two kinds of VLPs. Furthermore, BPV-VP2 VLPs were praised for their greater purity and integrity than BPV-VP1VP2 VLPs, as indicated by SDS‒PAGE. Therefore, our research demonstrates that the function of VP1 has no bearing on the stability or integrity of BPV-VLPs. CONCLUSIONS: In summary, incredible physiochemically stable BPV VP2-derived VLPs have been found to be promising candidates for the development of multivalent vaccines and immunodiagnostic kits against enteric viruses and to carry heterogeneous epitopes for various economically important livestock diseases.

Delay in seeking treatment and associated factors among pulmonary tuberculosis patients attending public health facilities in the Metekel zone, Benishangul Gumuz region, Western Ethiopia.

Background: Tuberculosis is a major global public health problem and a leading cause of morbidity and mortality in Ethiopia. TB prevention and control in low-income countries, such as Ethiopia, face significant challenges, including late detection and treatment initiation. A delay in the initiation of tuberculosis treatment increases the morbidity and mortality of patients and community transmission. Therefore, this study aimed to assess patient delay and associated factors among pulmonary tuberculosis patients attending public health facilities in the Metekel Zone, Benishangul Gumuz Region, Western Ethiopia. Methods: An institution-based cross-sectional study was conducted from March to August 2020 among newly diagnosed pulmonary tuberculosis patients. All pulmonary tuberculosis patients (416) who came to all public health facilities of the Metekel zone for treatment during the period were included. Data were collected through face-to-face interviews using a structured and pretested questionnaire. A multivariable logistic regression was fitted to identify independent factors for delay in seeking treatment among PTB patients. Adjusted odds ratios with 95% CIs were determined, and variables with p values <0.05 were considered statistically significant. Results: Nearly three-fourths 302 (72.6, 95% CI: 68.5, 76.7) of the patients were delayed in seeking medical advice, with a median patient delay of 27 days (IQR: 21-31). Age of the patients [above 54 years (AOR = 2.65, 95% CI: 1.30, 5.40), 36-54 years (AOR = 1.86, 95% CI: 1.14,3.02)], family size of 5 members and above (AOR = 1.62, 95% CI: 1.10-3.14), travel time above 60 min (AOR = 3.65, 95% CI: 1.55, 8.60), history of visits to informal care providers (AOR = 1.74, 95% CI: 1.11, 3.14), and poor knowledge about PTB (AOR = 1.64, 95% CI: 1.04-2.44) were statistically significant factors associated with delays in seeking treatment among PTB patients. Conclusion: Most pulmonary tuberculosis patients delay seeking medical advice for their illnesses. Delays in seeking treatment were associated with older age, large household size, longer travel time to reach the nearby health facility, visiting informal care providers, and poor knowledge about pulmonary tuberculosis. Hence, it is crucial to consider community screening programs, enhance public awareness, and ensure the accessibility of TB diagnostic and treatment services.

Evaluation of the Emergency Obstetric and Newborn Care training in Gondar, Ethiopia; a mixed methods study.

In Ethiopia maternal and perinatal morbidity and mortality remains high. Timely access to quality emergency obstetric and neonatal care is essential for the prevention of adverse outcomes. Training healthcare providers can play an important role in improving quality of care, thereby reducing maternal and perinatal mortality and morbidity. The aim of this study was to evaluate change of knowledge, skills and behaviour in health workers who attended a postgraduate Emergency Obstetric and Newborn Care training in Gondar, Ethiopia. A descriptive study with before-after approach, using a mix of quantitative and qualitative data, based on Kirkpatrick's model for training evaluation was conducted. The evaluation focussed on reaction, knowledge, skills, and change in behaviour in clinical practice of health care providers and facilitator's perspectives on performance. A 'lessons learned approach' was included to summarize facilitators' perspectives. Health care providers reacted positively to the Emergency Obstetric and Newborn Care training with significant improvement in knowledge and skills. Of the 56 participants who attended the training, 44 (79%) were midwives. The main evaluation score for lectures was 4,51 (SD 0,19) and for breakout sessions was 4,52 (SD 0.18) on scale of 1-5. There was a statistically significant difference in the pre and post knowledge (n = 28, mean difference 13.8%, SD 13.5, t = 6.216, p<0.001) and skills assessments (n = 23, mean difference 27.4%, SD 22.1%, t = 5.941, p<0.001). The results were the same for every component of the skills and knowledge assessment. Overall, they felt more confident in performing skills after being trained. Local sustainability, participant commitment and local context were identified as challenging factors after introducing a new training program. In Gondar Ethiopia, the Emergency Obstetric and Newborn Care training has the potential to increase skilled attendance at birth and improve quality of care, both vital to the reduction of maternal and perinatal mortality and morbidity.

Identification of serotypes of Mannheimia haemolytica and Pasteurella multocida from pneumonic cases of sheep and goats and their antimicrobial sensitivity profiles in Borana and Arsi zones, Ethiopia.

Respiratory diseases caused by Mannheimia haemolytica (M. haemolytica) and Pasteurella multocida (P. multocida) have been known to result in a considerable loss due to mortality and reduced production. This study aimed at isolation and identification of M. haemolytica and P. multocida associated with pneumonic pasteurellosis in sheep and goats using bacteriological and molecular techniques. Identification of serotypes of M. haemolytica and P. multocida was done using indirect haemagglutination test. The in vitro antimicrobial sensitivity profiles of the M. haemolytica were tested using standard disk diffusion method. A total of 52 and 78 nasal swabs were collected from pneumonic cases for bacterial isolation and identification in Borana and Arsi zone, respectively. Four hundred sera samples were collected for identification of serotypes. The results showed that 17 of 52 (32.69%; 95% CI 20.33, 47.11) nasal swabs collected from pneumonic animals in Borana yielded positive results for Pasteurella/Mannheimia species, 13 (25.00%; 95% CI 14.03, 38.95) of which were M. haemolytica. None of the samples yielded P. multocida. Twenty-three of 78 (29.49%; 95% CI 19.69, 40.89) nasal swabs collected at Arsi from pneumonic animals yielded positive results for M. haemolytica (17) and P. multocida (6). Secondary biochemical characterization revealed that 14 of the 17 isolates conform to M. haemolytica whereas none of the 6 isolates suspected to be P. mutocida were confirmed. Eleven (84.62%) isolates from Borana and 4 (28.57%) from Arsi were confirmed to be M. haemolytica using PCR targeting the Rpt2 genes. Assay for M. haemolytica serotype A1 revealed all belong to A1. None of the isolates with cultural and morphological features of P. multocida gave positive results by molecular assay. Serological assay identified three serotypes of M. haemolytica namely A1, A2 and A7 almost in all of the samples whereas P. multocida serotype A was detected in 78.75% of the samples. The M. haemolytica isolates tested for susceptibility to antibiotics showed resistance against Bacitracin (83.33%) and Penicillin (50.00%) while they were found susceptible to Gentamycin (100%), Chloramphenicol (100%) and Sulfamethoxazole (100%) and Tetracycline (83.33%). In conclusion, the results of the present study revealed the association of M. haemolytica with pneumonic pasteurellosis in sheep and goats and can be of use in vaccine development in Ethiopia. Nevertheless, further investigations and continuous monitoring of antimicrobial resistance and appropriate selection and prudent use of antimicrobials in livestock sector are required.

Prioritization of neglected tropical zoonotic diseases: A one health perspective from Tigray region, Northern Ethiopia.

Neglected tropical zoonotic diseases (NTZDs) continue to have a major effect on the health of humans and animals. In this study, a one health approach was used to prioritize and rank neglected tropical zoonotic diseases at the regional and zonal levels in Tigray National Regional State, Ethiopia. For prioritization of NTZDs a cross-sectional study through a structured questionnaire was administered to 313 health experts from human and animal health sectors. In addition, focus group discussions (FGD) were held with purposively selected key informants. Descriptive, and Multivariable analysis was applied to report the results and a ranked list of diseases was developed at the zonal and regional level. In the region, 8 of the 12 World Health Organization listed NTZDs were considered major diseases including anthrax, brucellosis, bovine tuberculosis, taeniasis, leishmaniasis, rabies, schistosomiasis, and soil-transmitted helminths. Considering the zoonotic and socioeconomic importance of the diseases at the regional level, rabies ranked 1stwhereas anthrax, bovine tuberculosis, leishmaniasis, and brucellosis were ranked from 2nd to 5th, respectively. The FGD result also supported the prioritization result. The Multivariable analysis showed a statistically significant difference in the zonal distribution of anthrax (р = 0.009, OR = 1.16), taeniasis (p<0.001, OR = 0.82), leishmaniasis (p<0.001, OR = 1.91), rabies (p = 0.020, OR = 0.79) and soil-transmitted helminths (p = 0.007, OR = 0.87) but not for brucellosis (p = 0.585), bovine tuberculosis (p = 0.505), and schistosomiasis (p = 0.421). Anthrax (p<0.001, OR = 26.68), brucellosis (p<0.001, OR = 13.18), and taeniasis (p<0.001, OR = 6.17) were considered as the major zoonotic diseases by veterinary practitioners than human health practitioners whereas, leishmaniasis was recognized as a major health challenge by human health professionals. Understanding the priority diseases in the region is supportive for informed decision-making and prioritizes the limited resources to use. Furthermore, strengthening the collaboration between human and animal health professions is important to control the diseases.

Knowledge of obstetric danger signs and associated factors among mothers in Bahir Dar district, northwest Ethiopia: an institution-based cross-sectional study.

BACKGROUND: Like other developing countries, in Ethiopia, obstetric complications contribute to about 50% of the maternal deaths. Thus, the aim of this study was to assess knowledge of obstetric danger signs and its associated factors among mothers attending the postnatal clinic at Felege Hiwot Referral Hospital, Bahir Dar district, northwest Ethiopia. METHODS: A hospital-based cross-sectional study was conducted on 410 postnatal mothers at Felege Hiwot Referral Hospital from June to September 2015. Knowledge of obstetric danger signs among postnatal clinic attending mothers was determined by using seventeen obstetric danger sign questions via interviewing. The binary logistic regression model was used to identify associated factors. In the multivariable analysis, variables with a P value of < 0.05 were considered statistically significant. Odds ratio with 95% confidence interval (CI) was calculated to show the strength of association. RESULTS: About 59% [95% CI 55, 63] of mothers were knowledgeable about obstetric danger signs. The odds of having good knowledge of obstetric danger signs were higher among mothers who were more educated [AOR = 6.86, 95% CI 2.47, 19.27], earned more than 3500 ETB household monthly income [AOR = 3.38, 95% CI 1.20, 13.96], and received information on danger signs from health extension workers (HEWs) [AOR = 4.23, 95% CI 1.83, 9.70] compared to their counterparts. However, mothers with service utilization decision power [AOR = 0.14, 95% CI 0.07, 0.27] with husband were 86% times less likely to be knowledgeable compared to mothers decided by themselves. CONCLUSION: In this study, below two thirds of mothers had good knowledge of obstetric danger signs. Thus, improving educational status and obtaining health information on obstetric danger signs from health professionals should be intensified. This implies that lack of awareness may lead to delay in seeking care. Thus, improving mothers' socio-economic status and self-decision-making power on utilization of health service are essential to mitigate the high burden of maternal morbidity due to obstetric complications. Also, obtaining health information on obstetric danger signs from health professionals should be intensified.

Rubella virus infections and immune status among pregnant women before the introduction of rubella vaccine in Amhara Regional State, Ethiopia.

BACKGROUND: Rubella and its associated congenital anomalies have been greatly reduced in most developed countries through use of the rubella vaccine. However, the magnitude of the problem is underestimated and there are no well-established rubella/congenital rubella syndrome prevention and control strategies in many developing countries, including Ethiopia. The aim of this study was to determine the prevalence of rubella virus infections among pregnant women and their immune status before the introduction of rubella vaccine in Amhara Regional State, Ethiopia. METHODS: A prospective cross-sectional study was conducted among pregnant women in Dessie, Felege-Hiwot, and University of Gondar referral hospitals, from December 2015 to February 2017. After obtaining written informed consent, socio-demographic data, reproductive history, clinical manifestations, and the possible risk factors for rubella virus infections were collected using a structured questionnaire. The laboratory analysis of rubella-specific antibodies was done using an enzyme-linked immunoassay method on venous blood samples. Data were entered and analyzed using IBM SPSS Statistics version 20. Binary logistic regression was used to determine the strength of association between the dependent variables and covariates. RESULTS: A total of 600 pregnant women were included in the study. Their mean age was 26.4±5years (range 16-40 years). The overall seroprevalence of rubella infection was 89%. Of the total study participants, 9.5% were positive for rubella-specific IgM antibody, which indicates acute/recent rubella virus infection. In contrast, 79.5% of them had protective levels of rubella-specific IgG antibody and were immune as a result of previous wild-type rubella infection. However, 11% of the pregnant women were negative for both rubella-specific antibodies; these women represent the susceptible group. CONCLUSIONS: A large number of pregnant women had acute/recent rubella virus infections at the time of data collection, indicating that the virus is endemic in the study area. More than a tenth of pregnant women were found to be susceptible to acquiring the infection in future pregnancies, with the possible risk of rubella-associated congenital anomalies. Hence screening of all women of child-bearing age before conception and during pregnancy might reduce the devastating effects of the virus on the developing fetus.

Prevalence, risk factors and antimicrobial susceptibility test of Staphylococcus aureus in Bovine cross breed mastitic milk in and around Asella town, Oromia regional state, southern Ethiopia.

A cross sectional study was undertaken from November 2016 to March 2017 in and around Asella town, Oromia regional state, southern Ethiopia, to determine the prevalence, associated risk factors and antimicrobial susceptibility of Staphylococcus aureus, in Bovine cross breed mastitis milk. A total of 384 lactating dairy cows were screened for mastitis based on clinical examinations and California mastitis test (CMT). Out of 230 lactating crossbred cows with either clinical or subclinical mastitis examined for the involvement of Staphylococcus aureus. Staphylococcus aureus was isolated at a rate of 47.2% (N=92) and 42.9% (N=15) of the sub- clinical and clinical cases, respectively. The overall prevalence of Staphylococcus aureus scored in this study was 46.5% (N=107). Descriptive statistics and chi-square were used in order to assess the magnitude of the difference of comparable variables, as a result, among risk factors considered, Age, parity, and lactation stage were found significantly associated with the occurrence of S. aureus in mastitis milk (p<0.05). The current study revealed that S. aureus has 0% susceptibility to penicillinG, followed by tetracycline (14.2%). However, these randomly selected isolates were found to be totally (100%) susceptible to the Kanamycin. The possible justification for, low antimicrobial susceptibility to these commonly used antimicrobials might be repeated and uncontrolled use of these drugs without veterinarian's prescription. Proper provention and regular antimicrobial sensitivity testing helps to select effective antibiotics and ultimately reduce the development of resistance towards commonly used antibiotics. To conclude, the study was able to show that, mastitis caused by S. aureus is one of the major problems of dairy cows in milk production and imposing public health hazard in study area. Hence, every possible control and prevention strategies should be implemented.

Prevalence of salivary gland hypertrophy syndrome in laboratory colonies and wild flies of Glossina pallidipes in Ethiopia.

Glossina pallidipes salivary gland hyperplasia (GpSGH) syndrome caused by the salivary gland hyperplasia virus reduces the reproduction potential of tsetse flies, posing a serious threat for rearing of sufficient colonies for use of tsetse and trypanosome control using the sterile insect technique. This research was conducted in the Kaliti Tsetse Mass Rearing and Irradiation Centre in Ethiopia with the objective of studying the prevalence of GpSGH syndrome in laboratory colonies of G. pallidipes (Tororo and Arbaminch) reared for release in the implementation of the sterile insect technique and a field strain of G. pallidipes Arbaminch. Presence or absence of GpSGH was determined when pathological features of the salivary gland were revealed after dissection. The overall prevalence of GpSGH syndrome in laboratory colonies was 48.3% (747/1548) with a statistically significant (z = 17.30, p = 0.001) prevalence of 70.2% (544/775) in Arbaminch colonies and 26.26% (203/773) in Tororo colonies. The prevalence of GpSGH in laboratory flies fed according to the clean blood feeding protocol was 68.9% and 22.4% in Arbaminch and Tororo strains respectively. It was 70.5% and 27.2% respectively in laboratory colonies of Arbaminch and Tororo strains fed according to the standard membrane feeding protocol. The difference in prevalence of the disease between the two feeding protocols was not statistically significant in either Arbaminch (z = 0.361, p = 0.359) or Tororo (z = 1.22, p = 0.111) strains. The prevalence of SGH in wild G. pallidipes Arbaminch strain was 3% (15/500) and was significantly (z = 23.61, p < 0.001) lower than in the laboratory strain. The effect of age and density-related stress on the development of GpSGH was not statistically significant. The prevalence of GpSGH in the newly emerging (teneral) flies in the laboratory colonies was 66.7% and 20% in the Arbaminch and Tororo strains respectively. For all considered risk factors, the prevalence was much higher in G. pallidipes Arbaminch laboratory colonies.

Bovine tuberculosis and brucellosis prevalence in cattle from selected milk cooperatives in Arsi zone, Oromia region, Ethiopia.

BACKGROUND: Bovine tuberculosis (BTB) and bovine brucellosis are two important milk-borne zoonoses that have been shown to be prevalent to various degrees in Ethiopian cattle. The study was carried out in four Woredas (districts) around Asella town, Arsi Zone between October 2011 and March 2012 and included 318 small-holders in 13 dairy cooperatives that marketed the delivered milk. The aims of the study were i) to assess the prevalence of the two diseases in cattle in a cross-sectional study, ii) to assess potential risk factors of BTB and brucellosis to humans as well as the knowledge-attitude-practice (KAP) among these farmers towards these diseases. RESULTS: BTB testing using the comparative intradermal skin test (CIDT) was done on 584 milking cows, out of which 417 were serologically tested for brucellosis using the Rose Bengal Plate Test and reactors confirmed with an indirect ELISA test (PrioCHECK®). The individual animal prevalence was 0.3% (95% CI 0.1% to 1.3%) for BTB, 1.7% (95% CI 0.8% to 3.5%) for brucellosis and 8.9% (95% CI 6.8% to 11.5%) for MAC (Mycobacterium avium complex). Of the 13 milk cooperatives, two had at least one positive BTB reactor and five had animals positive for brucellosis. Cross-breeds accounted for 100% and 71.4% of the BTB and brucellosis reactors respectively. For both diseases, there were prevalence variations depending on Woreda. No animal was concomitant reactor for BTB and brucellosis. Raw milk was consumed by 55.4% of the respondents. 79.2% of the respondents reported touching the afterbirth with bare hands. The latter was fed to dogs in 83% of the households. One cow among the herds of the 130 interviewees had aborted in the last 12 months. Among the interviewees, 77% stated knowing tuberculosis in general but 42 out of the 130 respondents (32.3%) did not know that BTB was transmitted by livestock. Less than half (47.7%) of the respondents knew about brucellosis. CONCLUSIONS: Low prevalence of both diseases reflected the potential for the area to compete with the growing milk demand. The authors discussed the possible control strategies for the area.